Monocrotaline-induced structural remodeling of the intra-acinar pulmonary arteries and pulmonary hypertension

Summary The monocrofaline-induced structural changes of small pulmonary arteries in rat and their relationship with pulmonary hypertension and right ventricular hypertrophy were observed by determining the right ventricular systolic pressure, and by light and electron microscope and morphometry. One to 38 days after last injection of monocrotaline (MCT), a medial thickening and lumen marrrowing of the circular muscular arteries (CMA), accompanying terminal (TB) and respiratory bronchioles (RB), were found. And there after the lumen of CMA, accompanying TB, became dilated, and its medial thickness (MT) decreased, whereas the histopathologic changes of the partially muscular arteries (PMA), accompanying RB, became severe, their MT increased continuously, and finally reached the peak value on Day 50. At the first day after last MCT treatment, inflammation and muscularization were found in PMA and nonmuscular arteries (NMA), and became more severe with the cause of disease. Therefore, the intra-acinar pulmonary arteries, both CMA and PMA, increased in number while the NMA decreased in number significantly because of the structural remodeling. Four days after MCT treatment, the right ventricular systolic pressure began to rise, and reached its peak value on Day 50. Eight days after MCT injection, right ventricular hypertrophy developed, and became most significant from Day 23 to Day 30. The results suggest that structural remodeling, i.e. muscularization, of intra-acinar pulmonary arteries plays an important role in the development of pulmonary hypertension and right ventricular hypertrophy.     

应用改进的AdEasy系统制备重组腺病毒

目的 改进AdEasy系统，使之更简便、高效、快捷。方法 pAdtrack经酶切线性化后，转化经氯化钙处理的感受态AdEasier-1细菌，获得重组腺病毒质粒。最后将线性化的重组腺病毒质粒转染293细胞包装获得重组腺病毒。荧光显微镜下细胞中重组腺病毒观察绿色荧光蛋白的表达。结果 应用氯化钙法由pAdtrack转化AdEasier-1细菌，可获得极高比例的(20／20)阳性重组腺病毒克隆。结论 应用改进后的AdEasy系统构建腺病毒相比于传统AdEasy系统，方法更简便、成功率更高、实验周期更短、对实验室的条件要求低。     

Preparation and identification of monoclonal antibodies against Penicillium marneffei]

OBJECTIVE: To prepare and identify monoclonal antibodies (mAbs) against Penicillium marneffei. METHODS: Recombinant mannoprotein1 (MP1) of Penicillium marneffei was used to immunize BALB/c mice, and anti-MP1 mAbs were obtained by means of hybridoma. Screening and identification of the mAbs were subsequently performed with indirect enzyme-linked immunosorbent assay and Western blotting, respectively. RESULTS: Four hybridomas producing antibodies against Penicillium marneffei were obtained, and the IgG isotypes of the 4 mAbs were identified as IgG1 with affinity constants (K) of 8.2x10(-9), 4.7x10(-9), 6.5x10(-9) and 2.7x10(-9), respectively. Western blotting demonstrated specific recognition of Aspergillus fumigatus MP1 by the obtained mAbs. CONCLUSION: The 4 hybridomas producing anti-MP1 mAbs with high specificity and affinity can be of significant value in the diagnosis of Penicilliosis marneffei infections.     

The Inhibitory Effect of Radix Salviae Miltiorrhizae on Hypoxic Structural Remodeling of Intra－acinarPulmonary Arteries

(席思川)(车东媛)(张婉蓉)ＴｈｅＩｎｈｉｂｉｔｏｒｙＥｆｆｅｃｔｏｆＲａｄｉｘＳａｌｖｉａｅＭｉｌｔｉｏｒｒｈｉｚａｅｏｎＨｙｐｏｘｉｃＳｔｒｕｃｔｕｒａｌＲｅｍｏｄｅｌｉｎｇｏｆＩｎｔｒａ－ａｃｉｎａｒＰｕｌｍｏｎａｒｙＡｒｔｅｒｉｅｓ￥ＸＩＳｉ－ｃ...     

HBV、TB合并感染时抗TB治疗对肝功能的影响

目的:探讨乙型肝炎病毒(HBV)、结核(TB)杆菌合并感染时抗TB药物所致肝损伤的临床及组织学特点。方法:单纯TB感染者45例,TB合并HBV感染者35例,前4月均服用异烟肼、利福平、吡嗪酰胺、乙胺丁醇,后2月服用异烟肼、利福平。每2～4周复查肝功能、HBV标记物、T细胞亚群等指标,部分患者进行了肝穿刺病理检查。结果:TB HBV组肝功能异常发生率为34.3%,较单纯TB组高,发生时间较单纯TB组早半月,发生平均年龄小8岁;谷丙转氨酶水平、肝组织学炎症积分均较TB组高(P<0.05);肝功能异常者中80%为HBeAb阳性,T细胞免疫功能紊乱较重,CD4 细胞明显减少(P<0.05)。结论:HBV、TB合并感染时,抗TB药物的肝毒性增强,治疗过程中应及时复查肝功能。     

In vivo and in vitro anti-tumour response of selenium-protein polysaccharide extracted from rich selenium Agaricus blazei

In this study, the anti-tumour activity of selenium-protein polysaccharide (SPP), a water extract of the rich selenium Agaricus blazei, was tested both in vivo and in vitro. The results of in vivo experiments show that SPP at doses of 50 and 100 mg/kg inhibits proliferation of implanted Sarcoma 180 by 22 and 37.69%, respectively, and promotes lymphocyte transformation and natural killer (NK) cells activity in tumour bearing mice. During the in vitro experiment, we treated the tumour and non-tumour bearing mice with SPP, and prepared serum treated with SPP (Serum_SPP). The results show that Serum_SPP, whether from tumour or non-tumour bearing mice, significantly inhibits K562 cells proliferation and induces their apoptosis, and also significantly increases caspase-3 activity of K562 cells. However, the difference in anti-tumour activity of Serum_SPP between tumour and non-tumour bearing mice is significantly different (p<0.01). The results, according to the studies both in vivo and in vitro, imply that SPP extracted from rich selenium A. blazei can inhibit growth of implanted Sarcoma 180 and promote lymphocyte transformation and NK cells activity in vivo. Additionally, Serum_SPP can inhibit proliferation and cause apoptotic morphological changes and the fragmentation of internucleosomal DNA, and increase caspase-3 activity of K562 cells in vitro, which indicates that apoptosis of K562 cells induced by Serum_SPP may be related to up-regulation of caspase-3.     

PATHOGENESIS OF EXTRINSIC ALLERGIC ALVEOLITIS AND PULMONARY FIBTOSIS INDUCED BY STREPTOMYCES THERMOHYGROSCOPICUS

There were 48 strains of thermophilic actinomyces isolated from the specimens of mouldy hay and sputum of the patients suffering from farmer’s lung (FL). Streptomyces thermohygroscopicus (STHs),one strain of them, was used for this investigation. The microorganisms were injected into the lungs of rabbits and rats by thyrocrico – or tracheocentesis. The result showed that the pathological changes in the lungs including macrophage alveolitis, granuloma formation and diffuse interstitial were similar to that induced by other thermophilic actinomyces. IgG and C3 deposition in the lesions were also observed by immunofluorescence examination. Specific immunocomplexes in the sera of some animals were detected by ELISA with the STHs was possibly one of the pathogens responsible for FL in China’s countryside.     

Recombinant VP9-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Banna virus (genus Seadornavirus)

Banna virus (BAV, genus Seadornavirus, family Reoviridae) is an arbovirus suspected to be responsible for encephalitis in humans. Two genotypes of this virus are distinguishable: A (Chinese isolate, BAV-Ch) and B (Indonesian isolate, BAV-In6969) which exhibit only 41% amino-acid identity in the sequence of their VP9.The VP7 to VP12 of BAV-Ch and VP9 of BAV-In6969 were expressed in bacteria using pGEX-4T-2 vector. VP9 was chosen to establish an ELISA for BAV, based mainly on two observations: (i). VP9 is a major protein in virus-infected cells and is a capsid protein (ii). among all the proteins expressed, VP9 was obtained in high amount and showed the highest immuno-reactivity to anti-BAV ascitic fluid.The VP9s ELISA was evaluated in three populations: French blood donors and two populations (blood donors and patients with a neurological syndrome) from Malaysia, representing the region where the virus was isolated in the past.The specificity of this ELISA was >98%. In mice injected with live BAV, the assay detected IgG-antibody to BAV infection 21 days post-injection, which was confirmed by Western blot using BAV-infected cells.The VP9 ELISA permits to determine the sero-status of a population without special safety precautions and without any requirements to propagate the BAV. This test should be a useful tool for epidemiological survey of BAV.     

新生大鼠海马神经元原代培养方法的研究

目的 :研究和比较 3种不同培养神经元的方法 ,提供一种纯化效率和存活率均较高的培养技术。方法 :取新生SD大鼠的海马区 ,应用一般培养法、加阿糖胞苷法、加阿糖胞苷和神经生长因子法培养神经元 ,观察神经元的形态。用倒置显微镜观察和MTT比色分析检测神经元的存活率。用ABC免疫组化染色法 ,比较 3种不同培养方法在不同培养时间所获得神经元的纯度。结果 :加阿糖胞苷和神经生长因子培养组神经元生长良好 ,纯度和存活率均较高。结论 :加阿糖胞苷和神经生长因子培养神经元的方法具有简便可行 ,易于生长的优点 ,可作为神经元体外培养的良好模型     

低氧致鼠腺泡内肺动脉PDGF—B链蛋白表达的变化

目的：探讨血小板源性生长因子（ＰＤＧＦ）在低氧肺动脉高压血管构形重建发生中的作用。方法：应用免疫组化技术结合计算机图像分析，检测了低氧大鼠腺泡内肺动脉（ＩＡＰＡ）ＰＤＧＦ－Ｂ链蛋白表达水平。结果：常氧时，ＩＡＰＡ仅有ＰＤＧＦ－Ｂ链蛋白弱表达；低氧１天时，ＩＡＰＡ便有较强的ＰＤＧＦ－Ｂ链蛋白表达，定位于ＩＡＰＡ的内皮和中膜，低氧３天至１４天仅分布于中膜；低氧１、３、５、７、１４天各时间点ＰＤＧＦ－Ｂ链蛋白表达分别为常氧组的１．５３、１．５９、１．５６、１．６２和１．４２倍，差异有显著性（Ｐ＜０．０１）。结论：ＰＤＧＦ－Ｂ链蛋白可能参与了低氧肺动脉高压血管构形重建的发病过程